本文關(guān)鍵詞：淋球菌外膜蛋白NspA和耐輻射奇球菌SOD基因的克隆和在乳酸菌中表達(dá)的初步研究　出處：《四川大學(xué)》2006年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 奈瑟菌表面蛋白A 超氧化物歧化酶 乳酸乳球菌 克隆 蛋白質(zhì)表達(dá) pMG36e pVE5523

【摘要】：目的 運(yùn)用乳酸菌胞內(nèi)表達(dá)載體pMG36e和分泌表達(dá)載體pVE5523，構(gòu)建淋病奈瑟菌表面蛋白A(Neisseria surface protein A，NspA)和耐輻射奇球菌錳超氧化物歧化酶(Mn superoxide dismutase，Mn-SOD)兩種目的基因的表達(dá)重組子，將重組子經(jīng)電穿孔轉(zhuǎn)化乳酸乳球菌，初步研究乳酸乳球菌表達(dá)融合NspA和SOD的情況，，為篩選目的基因的乳酸菌表達(dá)載體提供依據(jù)和思路：為后續(xù)開發(fā)淋球菌外膜蛋白的乳酸菌雙功能活菌疫苗提供參考；為進(jìn)一步研究SOD的表達(dá)對(duì)乳酸菌生物學(xué)特性的影響、開發(fā)可直接食用型SOD產(chǎn)品或微生態(tài)制劑奠定基礎(chǔ)。 方法 本研究內(nèi)容分兩部分： 1．pMG36e-nspA和pMG36e-sod重組子的構(gòu)建和在乳酸菌中表達(dá) 1) 優(yōu)化CTAB法提取淋球菌和耐輻射奇球菌基因組的條件；設(shè)計(jì)引物，優(yōu)化PCR擴(kuò)增目的基因nspA和sod的條件；目的基因分別與乳酸菌胞內(nèi)表達(dá)載體pMG36e連接，構(gòu)建表達(dá)重組子pMG36e-nspA和pMG36e-sod，將表達(dá)重組子轉(zhuǎn)化E．coli DH5a，利用紅霉素抗性篩選陽性克隆，質(zhì)粒小量提取后，雙酶切和測序鑒定。
[Abstract]:Purpose Lactic acid bacteria intracellular expression vector pMG36e and secretory expression vector pVE5523 were used. Neisseria surface protein A was constructed from Neisseria gonorrhoeae. NspA) and manganese superoxide dismutase (mn superoxide dismutase Mn-SOD) were expressed as recombinant genes. The expression and fusion of NspA and SOD of Lactococcus lactis were preliminarily studied by electroporation of the recombinant plasmid into Lactococcus lactis. To provide the basis and train of thought for screening the lactic acid bacteria expression vector of the target gene: to provide the reference for the further development of the Lactobacillus bifunctional live vaccine of the outer membrane protein of Neisseria gonorrhoeae; In order to further study the effect of SOD expression on the biological characteristics of lactic acid bacteria, the development of direct edible SOD products or microecological preparations laid the foundation. Method This research is divided into two parts: 1. Construction and expression of pMG36e-nspA and pMG36e-sod recombinant in lactic acid bacteria 1) to optimize the conditions for extracting the genomes of Neisseria gonorrhoeae and Radiococci by CTAB method. Primers were designed to optimize the conditions for nspA and sod amplification by PCR. Objective to construct the recombinant pMG36e-nspA and pMG36e-sod, respectively, by ligating the genes with the intracellular expression vector pMG36e of lactic acid bacteria. The recombinant plasmid was transformed into E. coli DH 5a. The positive clones were screened by erythromycin resistance screening. The plasmid was extracted in a small amount and identified by double enzyme digestion and sequencing.
【學(xué)位授予單位】：四川大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類號(hào)】：R371

【引證文獻(xiàn)】

相關(guān)期刊論文 前1條

1 潘偉芹;于建寧;王公金;徐小波;于峰祥;李燕;;SQR基因在大腸桿菌和乳酸菌中的表達(dá)[J];江蘇農(nóng)業(yè)學(xué)報(bào);2012年01期

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